“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Note. This article will be published in a forthcoming issue of theInternational Journal of Sports Physiology and Performance. The
article appears here in its accepted, peer-reviewed form, as it wasprovided by the submitting author. It has not been copyedited,
proofread, or formatted by the publisher.

Section: Original Investigation
Article Title: Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping
Performance
Authors: Pietro Luigi Invernizzi1, Eloisa Limonta1, Andrea Riboli1, Andrea Bosio2, Raffaele
Scurati1, and Fabio Esposito1,3
Affiliations: 1Department of Biomedical Sciences for Health, University of Milan, Via G. Colombo
71, 201333 Milan, Italy. 2Human Performance Laboratory, Mapei Sport Centre, Via Don Minzoni 34,
21053 Castellanza (VA), Italy. 3Centre of Sport Medicine, Don Gnocchi Foundation, Via Capecelatro
66, 20148 Milan, Italy.

Journal: International Journal of Sports Physiology and Performance
Acceptance Date: August 1, 2015
©2015 Human Kinetics, Inc.

DOI: http://dx.doi.org/10.1123/ijspp.2014-0507

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Effects of acute carnosine and -alanine on isometric force and jumping performance1

Pietro Luigi Invernizzi, 1Eloisa Limonta, 1Andrea Riboli, 2Andrea Bosio, 1Raffaele Scurati,
Fabio Esposito

1,31Department of Biomedical Sciences for Health, University of Milan, Via G. Colombo 71, 201333
Milan, Italy.
2
Human Performance Laboratory, Mapei Sport Centre, Via Don Minzoni 34, 21053 Castellanza(VA), Italy.
3
Centre of Sport Medicine, Don Gnocchi Foundation, Via Capecelatro 66, 20148 Milan, Italy

Corresponding author:
Fabio Esposito
Department of Biomedical Sciences for Health
University of Milan
Via G. Colombo 71
20133 Milan, Italy
Phone: +39-02-5031 4649
Fax: +39-02-5031 4630
E-Mail: fabio.esposito@unimi.it
Running head: Muscle performance with carnosine
Abstract word count: 250
Text only word count: 3470
n. of figures: 2
n. of tables: 3

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Abstract
Purpose: To assess the effects of acute L-carnosine and β-alanine (Carn-BA) supplementation
on isometric and dynamic tasks. Methods: Twelve healthy participants performed knee
extensors maximal voluntary contractions (MVC) and countermovement jumps (CMJ) before
and after a fatiguing protocol (45s continuous CMJ). Isometric and dynamic tests were
performed four hours after Carn-BA (2g of L-carnosine and 2g of -alanine) or placebo (PLA)
ingestion, in random order. After the fatiguing protocol, blood lactate concentration ([La-]),general and muscular rate of perceived exertion (RPE), and muscular pain (after 24h and 48hfrom the end of the fatiguing protocol) were assessed. Results: During the fatiguing protocol,
significant decreases in jump height, and increases in contact time were found in both groups
from the 15th second onwards to the end of the fatiguing protocol. Average contact time andjump height were respectively lower (-7%; P=0.018) and higher (+6%; P=0.025) in Carn-BA
compared to PLA. After the fatiguing protocol, MVC decreased in both PLA and Carn-BA,
but it was higher in Carn-BA compared to PLA (+15%, P=0.012), while CMJ did not change.
Moreover, general RPE was lower and muscle pain at 24h was higher in Carn-BA comparedto PLA, whereas muscular RPE and [La-] did not differ between conditions. Conclusions:
Ingesting Carn-BA prior to exercise induced positive effects on MVC and CMJ after thefatiguing protocol and improved CMJ performance during the 45s continuous jumping effort,
even when acutely supplemented. Furthermore, Carn-BA reduced the general RPE andincreased muscular pain 24 h after the fatiguing task.
Keywords: countermovement jump, maximum voluntary contraction, EMG, RPE, muscularpain

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Introduction
Muscle performance decreases progressively during exercise because of central andperipheral fatigue.1 It’s becoming very popular among recreational and competitive athletes
the ingestion of different types of legal ergogenic aids to counteract the effects of fatigue. 2, 3
Although controversial,4, 5 chronic supplementation with carnosine together with ß-alanine(Carn-BA), the rate-limiting precursor in carnosine synthesis, 6 has been recently proposed toincrease skeletal muscle carnosine levels, 6-8 which can improve muscle contractility andreduce muscle fatigue. 8, 9 Among several hypotheses, Carn-BA effects have been suggested torely on its buffering action in the muscle 10 or on its role as a diffusible Ca2+/H+ exchanger,
which would increase cross-bridge formation and force output.11
Four weeks of L-carnosine or ß-alanine supplementation has been shown to increase
muscle carnosine levels up to about 66 % and 64 %, respectively. 12 An amelioration of muscle
contractility and exercise performance have been associated to these carnosine increased levels.
6, 8, 13, 14

While the consequences of chronic Carn-BA supplementation have been alreadyextensively investigated, little is known about the effects of acute Carn-BA administration.
Gardner et al. 15 argued that carnosine could be already available inside the muscle fibres few
hours after ingestion. Therefore, acute Carn-BA supplementation may be similarly effective inincreasing muscle performance. Consequently, athletes could benefit from its ingestion
immediately, avoid the nuisance of chronic supplementation, simplify consumption
procedures, and reduce the associated costs. Bellinger and coworkers found that acute ingestionof ß-alanine was not associated with improved cycling time trial performance. 16 However, theeffects of acute supplementation on high intensity anaerobic tasks yet have to be assessed.
Therefore, this study aimed at assessing the effects of acute Carn-BA supplementation
on maximum isometric strength and repeated vertical jumps. Should a higher isometric and“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

jumping performance in Carn-BA compared to PLA be retrieved after a fatiguing protocol,then Carn-BA effects may be claimed to be present also after its acute supplementation. The
effects of acute Carn-BA ingestion on rate of perceived effort and muscular pain after afatiguing protocol were also assessed.
Methods
Participants
Twelve healthy, physically active, male participants (age 23 ± 2 years; stature 1.79 ±0.09 m; body mass 75.8 ± 9.8 kg; mean ± standard deviation, SD) volunteered for the study.
They were all without any form of neuromuscular and skeletal diseases. The participants were
naive to the purpose of the experiments. After full explanation of the experimental design andprocedures, each participant gave written informed consent before engaging in the experiments.
The study was approved by the local Ethics Committee and was conducted in accordance withthe principles of the 1964 Declaration of Helsinki.
Experimental design
The study has a double blind, randomized, counterbalanced placebo design. All
participants reported to the laboratory on three different occasions. The first visit served forfamiliarization purpose, during which participants were accurately instructed with theexperimental setup and procedures. In the two following visits, each participant performed
static and dynamic tests with (Carn-BA) or without (placebo-PLA) supplement ingestion, in
randomized order.
Carn-BA supplementation involved tablets with a total amount of 2 g of L-carnosineand 2 g of ß-alanine, plus excipients. According to the main formulator of the product (Velleja
Research, Piacenza, Italy), the carnosine and β-alanine combination was proposed to maximize
the carnosine content within the muscle cells. 10, 17 Placebo tablets contained only excipients.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Carn-BA and PLA tablets were both manufactured by Procemsa Farmaceutici(Nichelino, Turin, Italy). The purity of the supplement was tested independently from theformulator of the product, with the aim to exclude the presence of performance-enhancing
compounds such as anabolic agents or stimulants.
Visits 2 and 3 were separated by at least two weeks, according to the reported time forcarnosine washout. 18 During this period, participants were asked not to change their usual
training program and nutrition. The Carn-BA or PLA supplementation was carried out indouble-blind fashion and was administered four hours before tests. Some evidences, indeed,
indicates that carnosine becomes available and ready to exert its function inside the muscle
fibres just after few hours from ingestion. 19
During visits 2 and 3, after maximum voluntary contraction (MVC) assessment of theknee extensor muscles of the dominant limb, maximum countermovement jump (CMJ)
performance was evaluated. Then, a fatiguing protocol consisting of 45 s of continuous CMJ
jumps was administered. After the fatiguing protocol, one MVC and one CMJ were performed
again to assess the effects of fatigue.
Participants were asked not to engage in physical activity over 24 hours prior to theexperiments. All tests were carried out in a climate-controlled laboratory (constant temperatureof 21 ± 1 °C and relative humidity of 50 ± 5 %), approximately at the same hour of the day.
Experimental procedures
Initially all participants completed a standardized warm-up which consisted of 10 minof running at a slow pace, two sets of five isometric submaximal contractions and three trialsof vertical jumps.
MVC assessment. After the warm-up, participants completed three trials of maximal
isometric effort with the dominant leg knee extensors, each lasting 3 s, with 4 min of rest in“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

between. The highest value obtained was considered as the maximum voluntary contraction(MVC).
During tests, participants seated on a knee-extensor ergometer with the knee joint at 90
degrees. The load cell attached to the ergometer was connected to a carbon shin pad positionedon the dominant leg of the participant by an inextensible steel wire. The position of the subject
on the ergometer and the placement of the shin pad on the leg was carefully standardized. To
isolate the knee extensor muscles contribution to the force output, limiting accessory muscles
involvement, participants were also secured to the ergometer with Velcro straps at the chest.
After the fatiguing protocol, a single MVC determination was immediately repeated.
CMJ. Before the fatiguing protocol and three minutes after the last MVC assessment,
participants performed three single CMJ on a force platform (4 Jump, Kiestler, Zurich, CH),
with a recovery time of 3 min between trials. Participants squatted down until the knees werebent at 90°. The knee angle was measured by an electrogoniometer (Biopac System Inc.,
Goleta, CA, USA) and the full knee extension was assumed to be 0°. Operators firmly
encouraged the participants to jump as high as possible. After the fatiguing protocol and after
MVC reassessment, one CMJ was repeated in the same way as before. Each selected jump was
analyzed using a custom-built software (Labview 7.1, National Instruments, Austin, TX, USA)
that detected automatically different hallmarks (eccentric and concentric phase onsets, leaving
and landing instants), and measured the variables of interest. The landing instant was detected
as the instant at which ground reaction force exceeded 50 N. During each jump selected for the
analysis, the following parameters were calculated: peak force (pF), contact time, and jumpheight.
Force and EMG signals. During MVC and jump tests, force output andelectromyographic (EMG) signal from the vastus lateralis and vastus medialis muscles wererecorded. The force output of the knee extensor muscles was recorded using a previously

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

calibrated load-cell (mod. SM-2000N, Interface, Scottsdale, UK). The force signal was filtered(bandwidth 2–64 Hz) and stored on a personal computer after A/D conversion (mod. UM150,
Biopac System, Goleta, USA) with a sampling frequency of 2048 Hz. The EMG probe waspositioned on the belly of the muscle, with the electrodes (4 silver bars with 1 mm diameterand 1 cm length, 1 cm interelectrode distance) perpendicular to the fibers major axis. The skinarea under the EMG electrodes was shaved, gently abraded with fine sand paper and carefullycleaned with ethyl alcohol and conductive gel, to achieve an inter-electrode impedance below
2000 . EMG was amplified (gain of 1000; mod. ASE16, LiSin, Turin, Italy; input impedance:
60 M; common-mode rejection ratio: >90 dB), filtered (bandwidth 10–500 Hz) and stored ona personal computer after A/D conversion (mod. UM150, Biopac System, Goleta, USA) at asampling rate of 2048 Hz.
To avoid interference of transient phenomena, only the middle 2 s of the MVC test,
where force signal remained steady, were analysed. From the EMG signal, time domain
analysis allowed the calculation of the root mean square (RMS) during MVC and CMJ. EMG
RMS values during CMJ were selected from the best jump, based on the following criteria: (a)the correct execution of the movement, determined by electrogoniometer (knee joint angle
reaching 90°) and operator’s visual inspection, and (b) the maximal vertical height achieved.
To account for daily variations, CMJ EMG RMS values were normalized to the highest EMG
signal collected during MVC maneuver.
Blood lactate concentration ([La-]). Arterialized blood samples (5 µl) were collectedfrom the ear lobe before tests and at minute 1, 3 and 6 of recovery after the fatiguing protocol.
Samples were immediately analysed using a portable system (Lactate Pro LT-1710, Arkray,
Japan) to determine [La-].
Fatiguing protocol. After 5 minutes of recovery from pre-fatigue tests, participants
performed 45 s of continuous CMJ, during which they verbally encouraged to jump with a“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

maximum frequency and height, always reaching a knee angle of 90° at the end of the eccentric
phase of the movement. Again, the knee angle was monitored by an electrogoniometer, withthe full knee extension assumed to be 0°.
Scales and surveys. The general and muscular RPE were measured immediately after
the fatiguing protocol using the Borg scale (category ratio, CR-10 20) to determine theperceptual level of effort. In addition to the information provided during the familiarization
visit, the same instructions were repeated to the participants at the beginning of theexperimental sessions. RPE scale was visible to the participants throughout the entire fatiguing
protocol. Immediately after, participants were requested to give a rating of their general andmuscular (lower limbs) perceived exertion. After being specifically instructed, participants
were asked to classify their muscular pain at the lower limbs 24 h and 48 h after the end of thefatiguing protocol on a visual analogic scale (VAS24h and VAS48h, respectively). In particular,
individuals were instructed to make a vertical mark on a 10 cm horizontal line, with the leftedge of the line representing “no pain at all”, and the right edge of the line representing the“worst pain imaginable”. 21
Statistics
Statistical analysis was performed with a commercially available software (SigmaStat,
Systat Software Inc., USA). A sample size of 12 participants was selected to ensure a statisticalpower >0.80. The normal distribution of the sampling was checked by a Shapiro-Wilk test. A
two-way (condition*time) ANOVA for repeated measures was utilized to detect statisticaldifferences during the R-CMJ test. A post-hoc Bonferroni test was applied when necessary toestablish the location of the differences. The magnitude of the changes were assessed using
effect size (ES) statistics with the Standard Error of ES Estimate and the ES lower and upper
95% confidence interval (CI). ES was classified as trivial for ES values < 0.2, small between
0.2-0.6, moderate between 0.6-1.2, large between 1.2-2.0, very large >2.0. 22 Statistical level

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

of significance was fixed at an α level < 0.05. Unless otherwise stated, results are expressed asmean ± SD.
Results
None of the participants reported the occurrence of side effects following the acuteingestion of Carn-BA, neither during the testing period nor in the next hours or the day after.
MVC pre- and post-fatiguing protocol
As shown in Table 1, MVC was not statistically different between conditions before
the fatiguing protocol (P=0.141, ES=-0.78, CI=-1.8 - 0.24). After the fatiguing protocol, MVC
decreased from baseline by ~19% and 12 %, in PLA and Carn-BA, respectively (P=0.001,
ES=2.23, CI=0.99 - 3.48 for PLA; P=0.001, ES=2.17, CI=0.93 - 3.40 for Carn-BA). However,
MVC after the fatiguing protocol was higher in Carn-BA compared to PLA (+15 %, P=0.012,
ES=-1.62, CI=-2.75 - -0.49). During MVC, EMG RMS was higher in PLA in respect to CarnBA, both before and after the fatiguing protocol (P=0.011, ES=-0.11, CI= -1.09 – 0.87 before
the fatiguing protocol; P=0.044, ES=-0.23, CI=-1.22 – 0.75 after the fatiguing protocol).
Moreover, EMG RMS values decreased with fatigue by 22 % (P=0.006, ES=1.01, CI=-0.03 –2.05) and by 20 % (P=0.003, ES=0.74, CI=-0.28 – 1.75) in PLA and in Carn-BA, respectively.
CMJ pre- and post-fatiguing protocol
During CMJ, no differences in EMG RMS between groups (PLA and Car-BA, P=0.212
and P=0.195 for before and after the fatiguing protocol, respectively) and conditions (before
and after the fatiguing protocol, P=0.069 and P=0.192 for PLA and Car-BA, respectively) wereretrieved.
Carn-BA did not improve CMJ height compared to PLA, neither before (P=0.652) nor
after (P=0.217) the fatiguing protocol. Nevertheless, Carn-BA prevented CMJ pF reduction

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

with fatigue. In PLA, indeed, the fatiguing protocol decreased CMJ pF by ~ 6 % (P=0.017,
ES=0.48, CI= -0.51 – 1.48), while in Carn-BA it did not (P=0.107).
Fatiguing protocol
As indicated in Table 2, Carn-BA had positive effects on the overall performance during
the fatiguing protocol. In Carn-BA, the mean contact time through the fatiguing protocol was
lower than in PLA by ~ 7 % (P=0.018, ES=0.42, CI= -0.57 – 1.41). The mean height of thejumps was higher in Carn-BA than in PLA by ~ 6 % (P=0.025, ES=-0.37, CI= -1.36 – 0.61).
During the fatiguing protocol, significant decreases in jump height (P<0.001 and P<0.001 in
Carn-BA and PLA, respectively; Figure 1, panel A), and increases in contact time (P<0.001
and P<0.001 in Carn-BA and PLA, respectively; Figure 1, panel B) were found in both groups
from the 15th second onwards to the end of the fatiguing protocol.
Carn-BA did not affect [La-] at the end of the fatiguing protocol (12.34 ± 3.46 and 11.91± 2.51 mM in PLA and Carn-BA, respectively; P=0.256).
ANOVA did not disclose significant differences in EMG RMS between Carn-BA andPLA during the fatiguing protocol, as shown in Figure 2.
Perceived exertion and pain
The general and muscular RPE and the level of muscular pain at 24 and 48 h after thefatiguing protocol in PLA and Carn-BA are provided in Table 3. Immediately after thefatiguing protocol, less general effort was perceived in Carn-BA than in PLA (-6 %; P=0.042,
ES=0.50, CI= -0.49 – 1.50). On the contrary, the muscular RPE measured at the end of thefatiguing protocol did not differ between PLA and Carn-BA (P=0.093). VAS administeredalong the two days following the experiment unveiled higher levels of muscular pain in CarnBA compared to PLA at 24 hours (+19 %, P=0.039, ES=-0.55, CI= -1.55 – 0.44), but similarvalues at 48 hours (P=0.472).

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Discussion
The main finding of the present study was that Carn-BA acute supplementation was
effective in ameliorating maximum static and dynamic tasks. With Carn-BA, the maximum
isometric force dropped to a lesser extent after a 45-s fatiguing protocol. Moreover, jumps
height was higher while contact time was lower throughout the fatiguing protocol, comparedto PLA. Lastly, despite muscle pain indices at 24 hours were higher with Carn-BA, muscular
RPE immediately at the end of the fatiguing protocol did not differ between conditions, andgeneral RPE was lower in Carn-BA. These data suggest that Carn-BA had positive effects on
static and dynamic performance during and after a high intensity fatiguing effort even whenacutely administered.
Acute effects of Carn-BA on MVC and jump performance before the fatiguing protocol
The influence of acute Carn-BA supplementation on the maximum isometric force andjump performance is a novel finding of this study. Our findings show a higher neuromuscular
activation with Carn-BA, suggesting the occurrence of phenomena at central levelneuromodulator. 23 However, they do not support the hypothesis of an acute effect of Carn-BA
on unfatigued muscle performance. Even though MVC and CMJ pF showed higher values in
Carn-BA with respect to PLA, the difference did not achieve statistical significance, suggestingthat the tendency of Carn-BA to ameliorate maximum static and dynamic performance needs
further investigation. In previous studies, the increase in muscle carnosine levels has beenshown to induce positive effects on muscle performance in continuous or intermittent exercise.
13, 24, 25

These observations, though, were made only after chronic supplementation. No studies

on the effects of Carn-BA on maximum isometric force output have been performed so far.
Therefore, comparisons with previous findings cannot be made.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Acute effects of Carn-BA on MVC and jump performance during and after the fatiguing
protocolAs expected, MVC decreased after the fatiguing protocol in both Carn-BA and PLA.
However, the MVC post-fatigue value in Carn-BA was higher than in PLA, suggesting thatmuscle contractile properties were less affected by the fatiguing protocol with acute Carn-BA
supplementation. Carn-BA effects on MVC were accompanied by similar effects on EMG
activity. In particular, the neural drive was higher in Carn-BA as before fatigue, andneuromuscular activation dropped to a lesser extent after the fatiguing protocol, suggesting thatafter the fatiguing protocol Carn-BA effects were evident also at the force output level.
Collectively, we cannot exclude that the higher MVC in Carn-BA after the fatiguing protocol
could be attributed to phenomena occurring at central level neuromodulator. 23 This
explanation, though, may require further investigation.
In addition to these Carn-BA positive effects on MVC, CMJ pF in Carn-BA did not
drop after the fatiguing protocol as in PLA (see Table 1), even though CMJ height did notchange with fatigue. A decrease in jump height would be reasonably expected after a fatiguing
jumping exercise. 26, 27 The discrepancy could be possibly explained by the higher number ofjumps performed in the two cited studies compared to those in the present investigation.
Moreover, the longer recovery time before performing the CMJ post-fatiguing protocol may
have played a role. Indeed, in our study the CMJ measurement was accomplished after MVC
assessment, as MVC was chosen as the main parameter to determine the presence of fatigue.
Consequently, MVC test was performed immediately at the end of the fatiguing protocol. This
experimental procedure delayed the CMJ test by about 6 minutes (for [La-] determination
during recovery), compared to less than 2 minutes from the end of the fatiguing protocol in thestudies of King and coworkers 26 and Skurvydas and coworkers. 27 This period likely allowed

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

the participants to recover from fatigue, at least in part, before maximum jump performancereassessment.
The positive effects of acute Carn-BA supplementation had urther support from thefindings during the fatiguing protocol (Table 2), where Carn-BA condition highlighted a lower
average contact time and a higher average jump height. Moreover, Carn-BA counteracted thefall in jump height and the rise in contact time after the first 15 seconds of the fatiguing
protocol, as shown in Figure 1. Compared to PLA, the better jump performance with Carn-BA
may be ascribed to the reported amelioration in buffering capacity and/or an increased Ca2+
unloading at the muscle level, where Carn-BA exerts its primary function. 6-8, 11, 17 Should theCarn-BA-induced increase in buffering capacity be the explanation, 19 muscle contractility may
have been considerably preserved in Carn-BA compared to PLA, thus limiting the drop injumping capacity. On the contrary, should the effect of Carn-BA on dynamic performance bebased on higher Ca2+ unloading at the sarcomere level,11 leading to an increase in cross-bridge
formation and force output, this would anyway explain the present findings. Further studies,though, involving muscle biopsies before and after the same fatiguing protocol would be
required to elucidate these or other explanations.
Noticeably, the different behavior in Carn-BA became evident after the first 10 secondsof the fatiguing protocol, leading to the hypothesis that the increased muscle carnosine content
exerted an effect few seconds from the beginning of the fatiguing exercise.
Acute effects of Carn-BA supplementation on RPE and pain
RPE and pain surveys showed an interesting and unexpected finding supporting thebeneficial effects of Carn-BA to improve the performance during and after a fatiguing exercise.
Indeed, RPE at the end of the fatiguing protocol should have been similar in PLA and CarnBA because of the exhaustive meaning of the fatiguing protocol and of the consequential
intensity constraint asked to the participants (all-out continuous jumps). However, while this is“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

what occurred for the muscular RPE, this was not the case for general RPE (lower value in
Carn-BA compared to PLA). A first explanation may come from the lower contact time inCarn-BA compared to PLA that may reflect a higher usage of the elastic component during thejumps, thus entailing a lower perception of effort. An alternative or complementary explanation
could be ascribed to some effects that carnosine has been reported to induce on the central
nervous system. 23 Carn-BA, indeed, can affect either the motor cortex or the generation of theeffort sensation through the corollary discharge, inducing a positive effect on the generalperception of effort. 28
Compared to PLA, muscular pain was higher in Carn-BA after 24 hours from thefatiguing protocol, but not different at 48 hours. This finding may help to further understandthe contribution of Carn-BA in improving muscle performance. Possibly, participants wereable to push harder with Carn-BA and stress their muscles to a higher extent, reporting betterjumps with the same muscular RPE as in PLA. However, they experienced more muscular pain
the day after the fatiguing exercise may be due to a higher delayed muscular soreness inducedby the greater effort in Carn-BA, returning to a level similar to PLA after 48 hours.
Conclusions
Acute Carn-BA supplementation few hours prior to exercise improved muscle
performance during and after a fatiguing effort of high-intensity. This acute supplementationinduced positive effects on MVC and CMJ after the fatiguing protocol and improved CMJ
performance during the 45 s continuous jumping effort. The increased muscular engagement
was accompanied by a reduction in general RPE, and led to a higher level of muscular pain 24
h after the fatiguing protocol. As a practical application, in sports involving high intensity
anaerobic efforts, athletes may take advantage of acute Carn-BA supplementation, which inthe present investigation reported benefits similar to other protocols based on chronic

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

supplementation. Further studies on Carn-BA dose-response and on the effects of different
diets on intracellular carnosine levels are required for a better understanding of the topic.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

References
1.

Noakes TD, St Clair Gibson A, Lambert EV. From catastrophe to complexity: a novel
model of integrative central neural regulation of effort and fatigue during exercise in
humans: summary and conclusions. Br J Sports Med. 2005;39(2):120-124.

2.

Gonzalez AM, Walsh AL, Ratamess NA, Kang J, Hoffman JR. Effect of a pre-workout
energy supplement on acute multi-joint resistance exercise. J Sports Sci Med.
2011;10(2):261-266.

3.

Spradley BD, Crowley KR, Tai CY, et al. Ingesting a pre-workout supplement
containing caffeine, B-vitamins, amino acids, creatine, and beta-alanine before exercise
delays fatigue while improving reaction time and muscular endurance. Nutr Metab(Lond). 2012;9:28.

4.

Ducker KJ, Dawson B, Wallman KE. Effect of Beta alanine and sodium bicarbonate
supplementation on repeated-sprint performance. J Strength Cond Res.
2013;27(12):3450-3460.

5.

Jagim AR, Wright GA, Brice AG, Doberstein ST. Effects of beta-alanine
supplementation on sprint endurance. J Strength Cond Res. 2013;27(2):526-532.

6.

Derave W, Ozdemir MS, Harris RC, et al. beta-Alanine supplementation augments
muscle carnosine content and attenuates fatigue during repeated isokinetic contraction
bouts in trained sprinters. J Appl Physiol. 2007;103(5):1736-1743.

7.

Suzuki Y, Ito O, Mukai N, Takahashi H, Takamatsu K. High level of skeletal muscle
carnosine contributes to the latter half of exercise performance during 30-s maximal
cycle ergometer sprinting. Jpn J Physiol. 2002;52(2):199-205.

8.

Baguet A, Bourgois J, Vanhee L, Achten E, Derave W. Important role of muscle
carnosine in rowing performance. J Appl Physiol. 2010;109(4):1096-1101.

9.

Smith AE, Moon JR, Kendall KL, et al. The effects of beta-alanine supplementationand high-intensity interval training on neuromuscular fatigue and muscle function. Eur
J Appl Physiol. 2009;105(3):357-363.

10.

Baguet A, Koppo K, Pottier A, Derave W. Beta-alanine supplementation reduces
acidosis but not oxygen uptake response during high-intensity cycling exercise. Eur J
Appl Physiol. 2010;108(3):495-503.

11.

Blancquaert L, Everaert I, Derave W. Beta-alanine supplementation, muscle carnosine
and exercise performance. Cur Opin Clin Nutr Metab Care. 2015;18(1):63-70.

12.

Harris RC, Tallon MJ, Dunnett M, et al. The absorption of orally supplied beta-alanine
and its effect on muscle carnosine synthesis in human vastus lateralis. Amino Acids.
2006;30(3):279-289.

13.

Hill CA, Harris RC, Kim HJ, et al. Influence of beta-alanine supplementation on
skeletal muscle carnosine concentrations and high intensity cycling capacity. Amino
Acids. 2007;32(2):225-233.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

14.

Suzuki Y, Ito O, Mukai N, Takahashi H, Takamatsu K. High level of skeletal muscle
carnosine contributes to the latter half of exercise performance during 30-s maximal
cycle ergometer sprinting. Jpn J Physiol. 2002;52(2):199-205.

15.

Gardner ML, Illingworth KM, Kelleher J, Wood D. Intestinal absorption of the intact
peptide carnosine in man, and comparison with intestinal permeability to lactulose. J
Physiol. 1991;439:411-422.

16.

Bellinger PM, Minahan CL. Performance effects of acute beta-alanine induced
paresthesia in competitive cyclists. Eur J Sport Sci. 2015:1-8.

17.

Artioli GG, Gualano B, Smith A, Stout J, Lancha AH, Jr. Role of beta-alanine
supplementation on muscle carnosine and exercise performance. Med Sci Sports Exerc.
2010;42(6):1162-1173.

18.

Suzuki Y, Nakao T, Maemura H, et al. Carnosine and anserine ingestion enhances
contribution of nonbicarbonate buffering. Med Sci Sports Exerc. 2006;38(2):334-338.

19.

Begum G, Cunliffe A, Leveritt M. Physiological role of carnosine in contracting
muscle. Int J Sport Nutr Exerc Metab. 2005;15(5):493-514.

20.

Borg E, Kaijser L. A comparison between three rating scales for perceived exertion andtwo different work tests. Scand J Med Sci Sports. 2006;16(1):57-69.

21.

O'Connor PJ, Cook DB. Exercise and pain: the neurobiology, measurement, andlaboratory study of pain in relation to exercise in humans. Exerc Sport Sci Rev.
1999;27:119-166.

22.

Batterham AM, Hopkins WG. Making meaningful inferences about magnitudes. Int J
Sports Physiol Perform. 2006;1(1):50-57.

23.

Boldyrev AA, Aldini G, Derave W. Physiology and pathophysiology of carnosine.
Physiol Rev. 2013;93(4):1803-1845.

24.

Culbertson JY, Kreider RB, Greenwood M, Cooke M. Effects of beta-alanine on muscle
carnosine and exercise performance: a review of the current literature. Nutrients.
2010;2(1):75-98.

25.

Hobson RM, Saunders B, Ball G, Harris RC, Sale C. Effects of beta-alanine
supplementation on exercise performance: a meta-analysis. Amino Acids.
2012;43(1):25-37.

26.

King T, Kaper G, Paradis S. Effects of Lower Extremity Anaerobic Fatigue on
Neuromuscular Function and Jumping Performance. J Exerc Physiol Online.
2013;16(4).

27.

Skurvydas A, Jascaninas J, Zachovajevas P. Changes in height of jump, maximal
voluntary contraction force and low-frequency fatigue after 100 intermittent orcontinuous jumps with maximal intensity. Acta Physiol Scand. 2000;169(1):55-62.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

28.

Marcora S. Perception of effort during exercise is independent of afferent feedback
from skeletal muscles, heart, and lungs. J Appl Physiol. 2009;106(6):2060-2062.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Figure 1: Jump height (panel A) and contact time (panel B) during the 45-s fatiguing
protocol with (Carn-BA) and without (PLA) supplementation. Error bars indicate standard
error values. *P<0.05 between conditions.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Figure 2: EMG root mean square (RMS) values, normalized to the value obtained during
MVC, throughout the 45-s fatiguing protocol with (Carn-BA) and without (PLA)
supplementation. Error bars indicate standard error values.

“Effects of Acute Carnosine and -Alanine on Isometric Force and Jumping Performance” by Invernizzi PL et al.
International Journal of Sports Physiology and Performance© 2015 Human Kinetics, Inc.

Table 1
PLA

Carn-BA

before687 (61)after
553 (52)b

before727 (31)after
637 (46)a, b

EMG RMS (mV)

0.355 (0.079)

0.278 (0.073)b

0.372 (0.088)a

0.298 (0.087)a,b

CMJ pF (N)

2041 (257)1916 (216)b

2125 (224)2045 (171)

CMJ height (cm)32.3 (5.9)

33.4 (7.3)

31.6 (7.3)

33.2 (6.5)

EMG RMS (mV)

0.216 (0.119)

0.210 (0.155)

0.220 (0.097)

0.204 (0.079)

MVC (N)

Static and dynamic tests results in the two conditions, before and after the fatiguing protocol.
MVC, maximum voluntary contraction; EMG RMS, EMG root mean square (during both
MVC and countermovement jump, CMJ); pF, force peak. Standard deviation values are given
in parentheses. PLA, placebo; Carn-BA, carnosine with -alanine supplementation. aP<0.05 vs
PLA; bP<0.05 vs before.

Table 2
Jumps (n.)

PLA39.5 (4.0)

Carn-BA
39.9 (3.8)

Average contact time (ms)647.1 (111.9)

604.7 (81.4)*

Average jumps height (cm)25.7 (3.1)

27.2 (3.8)*

Mean values along the 45 s rebound countermovement jump. Measurements’ standard
deviation is given in parentheses. *P < 0.05 vs PLA.

Table 3General RPE (au)PLA
8.0 (0.8)

Carn-BA
7.5 (1.0)*

Muscular RPE (au)

7.5 (1.6)

7.1 (1.5)

VAS24h (cm)

3.7 (1.1)

4.4 (1.3)*

VAS48h (cm)

2.2 (1.7)

2.1 (0.7)

Rate of perceived exertion (RPE) and pain at the end and after 24 and 48 hours from the endof the fatiguing task. Standard deviation values are given in parentheses. VAS, visual analogic
scale. *P<0.05 vs PLA.